Nucleosidic phosphorylating agent and methods
AssigneeGenentech, Inc.
InventorRoberto Crea
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Abstract
Compounds of the formula ##STR1## where R is a base labile substituted-phenyloxy protecting group, are useful as phosphorylating reagents in the synthesis of oligonucleotides by a phosphotriester method. Such compounds are made by the reaction of .beta.-cyanoethanol and a corresponding R-phosphorodichloridate. A method of phosphorylating involves reacting such compounds, in the presence of base, with a nucleoside or a polynucleotide having a free 3' hydroxyl group, having the 5'--0 position protected by an organic group labile in acidic medium and having any phosphotriester linking groups protected by an organic group labile in strong basic medium.
Claims11
A method of phosphorylating a moiety of structure ##STR8## wherein B is adenyl, thymyl, guanyl or cytosyl and Q is selected from the group consisting of R.sub.1 O-, and ##STR9## wherein n is zero or an integer from 1 to about 18, R.sub.1 is an organic group labile in acidic medium and R.sub.2 is an organic group labile in basic medium which comprises reacting said moiety with a phosphorylating reagent of structure ##STR10## wherein R is a base labile phenyloxy protecting group, in the presence of a weak base to form the product ##STR11##
Dependencies are inferred from the claim text.